Enzyme-based flow injection analysis system for glutamine and glutamate in mammalian cell culture media
Enzyme-based flow injection analysis system for glutamine and glutamate in mammalian cell culture media
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DOI：
10.1006/abio.1998.3044
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发表时间：
1999-03-01
影响因子：
2.9
通讯作者：
Pittner, F
中科院分区：
文献类型：
--
作者：
Mayer, C;Frauer, A;Pittner, F
我们提出了一个如何注射分析系统设计的在线监测谷氨酸和谷氨酰胺的设置。这些氨基酸代表哺乳动物细胞培养中的主要能量来源。由谷氨酸脱氢酶和天冬氨酸氨基转移酶组成的循环测定产生与样品中谷氨酸浓度成正比的NADH。然后用荧光光度法测量NADH。谷氨酰胺通过转化为谷氨酸来测定，谷氨酸被进料到循环测定中。谷氨酰胺转化为谷氨酸是由天冬酰胺酶催化的。由于天冬酰胺酶与谷氨酰胺的反应性相对较高，且最适pH值与谷氨酸脱氢酶相似，因此使用天冬酰胺酶代替谷氨酰胺酶。将酶共价固定到活化的可控孔玻璃珠上，并整合到流动注射分析系统中。本文介绍了固定化酶的应用及固定化酶的技术装置。(C)北京：科学出版社.
We present the setup of a how injection analysis system designed for on-line monitoring of glutamate and glutamine. These amino acids represent a major energy source in mammalian cell culture. A cycling assay consisting of glutamate dehydrogenase and aspartate aminotransferase produces NADH proportional to the glutamate concentration in the sample. NADH is then measured spectrophotometrically. Glutamine is determined by conversion to glutamate which is fed into the cycling assay. The conversion of glutamine to glutamate is catalyzed by asparaginase. Asparaginase was used in place of glutaminase due to its relatively high reactivity with glutamine and a pH optimum similar to that of glutamate dehydrogenase. The enzymes were immobilized covalently to activated controlled pore glass beads and integrated into the how injection analysis system. The application of the immobilized enzymes and the technical setup are presented in this paper. (C) 1999 Academic Press.